Tsuchiya T, Ueda M, Ochiai H, Imajoh-Ohmi S, Kanegasaki S
Institute of Medical Science, University of Tokyo.
J Biochem. 1993 Mar;113(3):395-400. doi: 10.1093/oxfordjournals.jbchem.a124057.
The 5'-flanking region of the human erythropoietin (Epo) gene contains a 0.14-kb sequence that is conserved in the Epo gene from mouse and located within a promoter that is activated under hypoxic conditions such as anemia. Using a fragment containing this sequence in DNA mobility shift assays, we found that specific DNA-binding proteins were induced in mouse kidney nuclei under anemic hypoxia. Using synthetic double-stranded oligonucleotides that contain this sequence, the essential binding site was defined to be the -40 to -20 region upstream of the transcription initiation site in the human Epo gene. By DNA affinity chromatography using a column with the immobilized 5'-flanking sequence, two inducible binding proteins with apparent molecular masses of 55 and 45 kDa were identified in the nuclei of mouse kidney and liver under anemic hypoxia. These binding proteins were also induced during cobalt exposure.
人类促红细胞生成素（Epo）基因的5'侧翼区域包含一段0.14 kb的序列，该序列在小鼠的Epo基因中保守，且位于在诸如贫血等缺氧条件下被激活的启动子内。在DNA迁移率变动分析中使用包含该序列的片段，我们发现在贫血性缺氧条件下小鼠肾细胞核中可诱导产生特异性DNA结合蛋白。使用包含该序列的合成双链寡核苷酸，确定关键结合位点为人类Epo基因转录起始位点上游-40至-20区域。通过使用固定有5'侧翼序列的柱进行DNA亲和层析，在贫血性缺氧条件下从小鼠肾和肝细胞核中鉴定出两种表观分子量分别为55 kDa和45 kDa的可诱导结合蛋白。在钴暴露期间也可诱导产生这些结合蛋白。
